Allogeneic administration of fetal membrane-derived mesenchymal stem cells attenuates acute myocarditis in rats

We reported previously that the autologous administration of bone marrow-derived mesenchymal stem cells (BM-MSC) significantly attenuated myocardial dysfunction and injury in a rat model of acute myocarditis by stimulating angiogenesis and reducing inflammation. Because BM aspiration procedures are invasive and can yield low numbers of MSC after processing, we focused on fetal membranes (FMs) as an alternative source of MSC to provide a large number of cells. We investigated whether the allogeneic administration of FM-derived MSC (FM-MSC) attenuates myocardial injury and dysfunction in a rat myocarditis model. Experimental autoimmune myocarditis (EAM) was induced in male Lewis rats by injecting porcine cardiac myosin. Allogeneic FM-MSC obtained from major histocompatibility complex-mismatched ACI rats (5 × 10⁵ cells/animal) were injected intravenously into Lewis rats one week after myosin administration. At day 21, severe cardiac inflammation and deterioration of cardiac function were observed. The allogeneic administration of FM-MSC significantly attenuated inflammatory cell infiltration and monocyte chemoattractant protein 1 expression in the myocardium and improved cardiac function. In a T-lymphocyte proliferation assay, the proliferative response of splenic T lymphocytes was significantly lower in cells obtained from FM-MSC-treated EAM rats that reacted to myosin than in cells obtained from vehicle-treated rats with EAM. T-lymphocyte activation was significantly reduced by coculture with FM-MSC. The allogeneic administration of FM-MSC attenuated myocardial dysfunction and inflammation, and the host cell-mediated immune response was attenuated in a rat model of acute myocarditis. These results suggest that allogeneic administration of FM-MSC might provide a new therapeutic strategy for the treatment of acute myocarditis.     

Myeloid-specific SIRT1 Deletion Aggravates Hepatic Inflammation and Steatosis in High-fat Diet-fed Mice

Sirtuin 1 (SIRT1) is a mammalian NAD⁺-dependent protein deacetylase that regulates cellular metabolism and inflammatory response. The organ-specific deletion of SIRT1 induces local inflammation and insulin resistance in dietary and genetic obesity. Macrophage-mediated inflammation contributes to insulin resistance and metabolic syndrome, however, the macrophage-specific SIRT1 function in the context of obesity is largely unknown. C57/BL6 wild type (WT) or myeloid-specific SIRT1 knockout (KO) mice were fed a high-fat diet (HFD) or normal diet (ND) for 12 weeks. Metabolic parameters and markers of hepatic steatosis and inflammation in liver were compared in WT and KO mice. SIRT1 deletion enhanced HFD-induced changes on body and liver weight gain, and increased glucose and insulin resistance. In liver, SIRT1 deletion increased the acetylation, and enhanced HFD-induced nuclear translocation of nuclear factor kappa B (NF-κB), hepatic inflammation and macrophage infiltration. HFD-fed KO mice showed severe hepatic steatosis by activating lipogenic pathway through sterol regulatory element-binding protein 1 (SREBP-1), and hepatic fibrogenesis, as indicated by induction of connective tissue growth factor (CTGF), alpha-smooth muscle actin (α-SMA), and collagen secretion. Myeloid-specific deletion of SIRT1 stimulates obesity-induced inflammation and increases the risk of hepatic fibrosis. Targeted induction of macrophage SIRT1 may be a good therapy for alleviating inflammation-associated metabolic syndrome.     

HPSE和Syndecan-1与人类早期复发性自然流产关系

目的 探讨乙酰肝素酶(heparanase,HPSE)和蛋白聚糖1(syndecan-1)在人类早期复发性自然流产绒毛组织中的表达及两者与复发性自然流产的关系.方法 采用实时荧光定量反转录聚合酶链反应(RT-PCR)分别检测山西大同地区非意愿性正常妊娠者(35例)和早期复发性自然流产者(28例)绒毛组织中HPSE和syndecan-1的mRNA表达水平,分析探讨两者与自然流产关系.结果 HPSE mRNA在正常妊娠组表达的阳性率为85.71％(30/35),明显高于复发性自然流产组的57.14％(16/28),差异有统计学意义(P＜0.05);而在正常妊娠组syndecan-1 mRNA的阳性表达率为54.28％(19/35),明显低于复发性自然流产组的82.14％ (23/28),差异亦有统计学意义(P＜0.05);HPSE和syndecan-1蛋白表达呈负相关(r=-0.386,P＜0.01).结论 在复发性自然流产中HPSE的表达降低,导致降解细胞外基质能力下降,syndecan-1表达增高,影响胚泡植入,导致自然流产的发生.     

GPC3在卵巢透明细胞腺癌中的表达及临床意义

目的:探讨磷脂酰肌醇蛋白聚糖3(glypican-3,GPC3)在卵巢透明细胞腺癌中的表达及临床意义?方法:采用免疫组织化学方法分别检测83例卵巢癌组织(透明细胞腺癌45例?浆液性癌20例?黏液性癌18例)?20例浆液性囊腺瘤及20例正常卵巢组织中GPC3蛋白的表达情况,实时荧光定量PCR法检测38例卵巢癌组织(透明细胞腺癌20例?浆液性癌10例?黏液性癌8例)?5例浆液性囊腺瘤和5例正常卵巢组织中mRNA的表达情况,并分析GPC3与卵巢透明细胞腺癌临床病理学参数及预后的相关性?结果:①卵巢透明细胞腺癌中GPC3蛋白阳性表达率和mRNA表达量(82.2%;1.326 1 ± 0.493 6)均明显高于浆液性癌(20%;0.426 5 ± 0.029 4)?黏液性癌(5%;0.265 2 ± 0.103 9)?浆液性囊腺瘤(0%;0.141 3 ± 0.011 3)?正常卵巢组织(0%;0.291 4 ± 0.048 1),差异有统计学意义(P均 < 0.01);②GPC3蛋白阳性表达率及GPC3 mRNA表达量与卵巢透明细胞腺癌的临床分期有相关性,Ⅲ~Ⅳ期患者中的表达(100%;1.808 1 ± 0.265 7)显著高于Ⅰ~Ⅱ期患者(71.4%; 1.045 8 ± 0.403 6)(P值 < 0.01);③卵巢透明细胞腺癌铂类药物耐药患者GPC3蛋白阳性表达率及mRNA表达量(100%; 1.808 1 ± 0.265 7)均高于铂类药物敏感型患者(70.3%; 0.991 8 ± 0.330 3)(P < 0.05);④GPC3蛋白阳性表达率与卵巢透明细胞腺癌患者的预后相关,预后差的患者中阳性率及表达量高,差异有统计学意义(P < 0.05),Kaplan-Meier单因素生存分析亦显示GPC3蛋白为影响预后的因素(P=0.048);⑤卵巢透明细胞腺癌中GPC3蛋白阳性表达率和mRNA表达量与患者的淋巴结转移和远处器官转移无相关性(P > 0.05)?结论:GPC3在病理鉴别诊断中有重要意义,GPC3与卵巢透明细胞腺癌患者的临床分期和铂类耐药相关,提示其可能在卵巢透明细胞腺癌的发生发展起重要作用,有望成为卵巢透明细胞腺癌的预后指标和潜在治疗靶标?     

核医学检查受检者所受辐射剂量分析

目的 对核医学检查受检者所受辐射剂量进行测量和分析,以有效剂量表征受检者受到的辐射强度。方法 对核医学检查受检者进行分类,并测量计算所受放射性药物的辐射剂量,受检者所受计算机断层扫描(CT)辐射剂量,通过CT扫描参数和受检者信息等计算得到,上述两者相加换算得到受检者检查所受的有效剂量,并分析受检者所受辐射剂量的影响因素。结果 受检者正电子发射断层计算机成像(PET-CT)检查受到正电子放射性药物¹⁸F-氟代脱氧葡萄糖(¹⁸F-FDG)、¹⁸F-氟代胸苷(¹⁸F-FLT)、¹¹C-胆碱(¹¹C-choline)、¹¹C-蛋氨酸(¹¹C-MET)和¹¹C-乙酸盐(¹¹C-Ac)辐射所致有效剂量分别为(5.06±0.73)、(4.74±1.29)、(1.71±0.05)、(3.18±0.69)和(1.08±0.19)mSv;CT常规扫描辐射有效剂量为(8.80±0.58)mSv,若增加诊断CT扫描,接受的有效剂量可增大至27 mSv;单光子发射计算机断层成像(ECT)检查受到单光子放射性药物⁹⁹Tc^m-亚甲基二磷酸盐(⁹⁹Tc^m-MDP)、⁹⁹Tc^m-大颗粒聚合白蛋白(⁹⁹Tc^m-MAA)、⁹⁹Tc^m-二乙基三胺五乙酸(⁹⁹Tc^m-DTPA)、⁹⁹Tc^m-甲氧基异丁基异腈(⁹⁹Tc^m-MIBI)和⁹⁹Tc^m-焦磷酸盐(⁹⁹Tc^m-PYP)辐照所致的有效剂量分别为(4.63±0.01)、(1.71±0.01)、(1.18±0.01)、(7.19±0.03)和(4.18±0.01)mSv。结论 核医学检查受检者受到放射性药物辐射的有效剂量在1.08~7.19 mSv之间,PET-CT检查中CT所致有效剂量是8.80 mSv。     

类风湿性关节炎患者外周血HFLC检测的临床意义

目的探讨外周血高荧光强度淋巴细胞(HFLC)与类风湿性关节炎(RA)发作的关系。方法选取2013年3月至2014年7月在武警后勤学院附属医院就诊的RA患者91例,含活动期患者(ARA)39例,静止期患者(SRA)52例,并选择同期114例该院查体患者作为健康对照组(HC),采用Sysmex XE-5000检测外周血HFLC的两参数绝对计数(HFLC#)和占白细胞百分比(HFLC%),采用SIMENS BNII全自动免疫散射比浊仪检测C反应蛋白(CRP)、血清免疫球蛋白(IgA、IgG、IgM)、类风湿因子(RF)、抗链O(ASO)、补体(C3、C4)。结果 HC组[0.00(0.00~1.00)×109/L]和SRA组[0.00(0.00~1.00)×109/L]间HFLC差异无统计学意义(P0.05),但均明显低于ARA组[1.00(0.00~2.00)×109/L](P0.05)。ARA组HFLC%[0.10%(0.00%~0.22%)]明显高于HC组[0.00%(0.00%~0.14%)](P0.05)。ARA组中HFLC#与IgE、CRP为正相关(P0.05),HFLC%与RF正相关(P0.05)。HFLC#与HFLC%的ROC曲线下面积分别为0.637(P0.05)、0.632(P0.05),当HFLC%界值为0.14%时,灵敏度为46.15%,特异度为78.85%。结论 HFLC%与HFLC#可以反映RA的发作,虽然其诊断效能不高,但是由于这些参数获取经济、便捷,可以为RA发作的诊断提供新的观察指标,为指导下一步的检查和治疗提供帮助。     

荧光定量PCR检测梅毒螺旋体方法的建立及初步应用

目的采用二聚体蝎型探针技术建立一种高敏感性和特异性的检测梅毒螺旋体(TP)的荧光定量聚合酶链反应(PCR)方法。方法根据TP特异性外膜蛋白Gpd的基因序列设计引物和荧光探针,采用基因工程技术构建可用于TP定量的标准品,优化荧光定量PCR的反应体系和反应条件,建立检测TP的二聚体蝎型探针定量PCR。采用本研究建立方法和商品化荧光定量PCR试剂盒检测40例临床标本,对比分析检测结果的统计学差异。结果成功构建了TP重组质粒标准品和TP的二聚体蝎型探针定量PCR,该方法线性范围为101~108 copy/mL,灵敏度为10copy/mL,特异性和敏感性均为100.0%;二聚体蝎型探针定量PCR对疑似梅毒病例阳性检出率显著高于目前商品化Taqman荧光定量PCR试剂盒(82.5%vs 62.5%,P0.05)。结论成功建立了二聚体蝎型探针荧光定量PCR快速检测TP的方法,为临床上TP的早期诊断和防控奠定了基础。     

Peroxiredoxin2基因RNAi慢病毒载体构建及对SW480细胞增殖的影响

目的：构建Peroxiredoxin2（PRDX2）基因RNA干扰（RNAinterference,RNAi）慢病毒表达载体,探讨PRDX2基因干扰后对结直肠癌SW480细胞增殖的影响。方法设计、合成靶向PRDX2的RNA干扰的序列,构建pGC‐EGFP‐shPRDX2慢病毒载体并进行鉴定,同时应用qRT‐PCR和Westernblot方法观察转染的SW480结肠癌细胞PRDX2mRNA和蛋白表达的抑制效果,并通过MTT、平板克隆形成实验检测细胞增殖变化。结果成功构建PRDX2基因慢病毒载体并经测序证实;pGC‐EGFP‐shPRDX2可有效抑制结直肠癌SW480细胞PRDX2的表达,感染慢病毒的SW480细胞中PRDX2mRNA和蛋白表达水平明显降低（P＜0．05）;SW480细胞经PRDX2RNA干扰后其生长和增殖能力显著降低（P＜0．05）。结论PRDX2基因RNAi慢病毒表达载体在SW480细胞中表达稳定可靠,PRDX2基因干扰后有效抑制了结直肠癌SW480细胞的增殖和生长,为进一步探讨PRDX2在结直肠癌发生、发展及转移中的作用奠定基础。     

Immunogenicity and safety of an E. coli-produced bivalent human papillomavirus (type 16 and 18) vaccine: A randomized controlled phase 2 clinical trial

BackgroundThis study aimed to investigate the dosage, immunogenicity and safety profile of a novel human papillomavirus (HPV) types 16 and 18 bivalent vaccine produced by E. coli.MethodsThis randomized, double-blinded, controlled phase 2 trial enrolled women aged 18–25 years in China. Totally 1600 eligible participants were randomized to receive 90 μg, 60 μg, or 30 μg of the recombinant HPV 16/18 bivalent vaccine or the control hepatitis B vaccine on a 0, 1 and 6 month schedule. The designated doses are the combined micrograms of HPV16 and 18 VLPs with dose ratio of 2:1. The immunogenicity of the vaccines was assessed by measuring anti-HPV 16 and 18 neutralizing antibodies and total IgG antibodies. Safety of the vaccine was assessed.ResultsAll but one of the seronegative participants who received 3 doses of the HPV vaccines seroconverted at month 7 for anti-HPV 16/18 neutralizing antibodies and IgG antibodies. For HPV 16, the geometric mean titers (GMTs) of the neutralizing antibodies were similar between the 60 μg (GMT = 10,548) and 90 μg (GMT = 12,505) HPV vaccine groups and were significantly higher than those in the 30 μg (GMT = 7596) group. For HPV 18, the GMTs of the neutralizing antibodies were similar among the 3 groups. The HPV vaccine was well tolerated. No vaccine-associated serious adverse events were identified.ConclusionThe prokaryotic-expressed HPV vaccine is safe and immunogenic in women aged 18–25 years. The 60 μg dosage formulation was selected for further investigation for efficacy.Clinical trials registration: NCT01356823.     

卵巢上皮性肿瘤患者人乳头状瘤病毒感染及核转录因子表达与临床相关性研究

目的探讨人乳头状瘤病毒(HPV)感染和核转录因子(NF-кB)表达与卵巢上皮性肿瘤的关系,为临床治疗提供依据。方法选取2008年1月-2013年10月71例卵巢肿瘤患者的病理标本,检测卵巢上皮肿瘤组织HPV感染和NF-кB表达情况,分析与临床、病理的相关性。结果卵巢上皮癌患者HPV的阳性率要明显高于良性肿瘤患者,分别为36.73%、9.09%,差异有统计学意义(P<0.05);卵巢上皮癌患者NF-кBp65的阳性率较良性肿瘤患者明显升高,分别为69.39%、17.17%,差异有统计学意义;HPV感染在卵巢癌不同临床病理结果比较,差异无统计学意义;卵巢上皮癌患者的临床分期、病理分级及CA125值在不同等级NF-кB阳性率的比较,差异有统计学意义(P<0.05),病理分化程度越低、临床分期越晚及CA125值越高,NF-кB阳性率越高。结论 HPV感染和NF-кB的高表达与卵巢癌的发生有一定的关系,HPV感染患者应予积极治疗,以预防卵巢癌的发生。